( Reaffirmed 2004 ) IS : 6393 - 1987 Indian Standard SPECIFICATION FOR 4WENYLACETAMIDE ( First Revision ) -0. FOREWORD Committee responsible for the preparation of this specification decided to revise it fin order to incorporate the modified requirement of the material in view of its improved~quality which is currently being made available in the country. A requirement of absence of cyanide has been introduced. 0.4 For the purpose of deciding whether a particular requirement of this standard is complied with, the final value, observed or calculated, expressing the result of a test or analysis, shall be rounded off in accordance with IS : 2- 1960". The number of significant places retained in the rounded off value should be the same as that of the specified value in this standard. *Rules for rounding off numerical values ( revised ). 0.1 This Indian Standard ( First Revision ) was adopted by the Bureau of Indian Standards on 30 October 1987, after the draft finalized by the Organic Chemicals ( Miscellaneous ) Sectional Committee had been approved by the Petroleum, Coal and Related Products Division Council. 0.2 a-phenylacetamide, the amide of phenylacetic acid is also known as phenylacetamide a-toluamide. It is mainly used in the manufacture of penicillin G. 0.3 This specification was published in 1971 on the basis of data and information made available by Regional Research Laboratory, Hyderabad; Indian Drugs and Pharmaceuticals Ltd, Hyderabad; and Hindustan Antibiotics Ltd, Pune. The 1. SCOPE 1.1 This standard prescribes the requirements and the methods of sampling and test for a-phenylacetamide. 2. REQUIREMENTS 2.1 Description - The form of white crystalline soluble in rectified spirit slightly soluble in water, material shall be in the powder. It shall be freely ( see IS : 323-1959" ) but ether and benzene. name and his recognized b) Manufacturers' trade-mark, if any; c) Mass of the material in the containers; and d) Lot or batch number, in code or otherwise. 3.2.2 The containers may also be marked * *.s . the Stanaara MarK. with . 2.2 The material shall also comply with the requirements given in Table 1 when tested according to the methods prescribed in Appendix A. Reference to the relevant clauses of Appendix A is given in co1 4 of Table 1. 3. PACKING AND MARKING 3.1 Packing - The material shall be packed in cardboard boxes, bottles, fibre drums, multiwalled paper sacks or as agreed to between the purchaser land the supplier. 3.2 Marking 3.2.1 The containers shall be marked with the following information: a) Name of the material; *Specification for rectified spirit ( revised ). NOTE - The use of the Standard Mark is governed by the provisions of the Bureau of Indian Standards Act 1986, and the Rules and Regulations made thereunder. The Standard Mark on products covered by an Indian Standard conveys the assurance that they have been produced to comply with the requirements of that standard under a well defined system of inspection, testing and quality control which is devised and supervised by BIS and operated by the producer. Standard marked products are also continuously checked by BIS for conformity to that standard as a further safeguard. Details of conditions under which a licence for the use of the Standard Mark may be granted to manufacturers or producers may be obtained from the Bureau of Indian Standards. 4. SAMPLING 4.1 Representative samples of the -material shalt be drawn and their conformity to the requirements of this standard be judged as prescribed in Appendix B. 1 IS: 6393-1987 TABLE 1 REQUIREMENTS FOR a-PHENYLACETAMIDE ( Clause 2.2 ) SL No. (0 i) ii) iii) iv) v) vi) vii) viii) ix) CHARACTERISTIC REQUIREMENT METHODOF TEST ( REP TO CL No. IN APPENDIX A ) (4) A-2 A-3 A-4 A-5 A-6 A-7 A-8 A-9 A-10 (2) Melting point, Min Loss on drying, percent Ammonium by mass, MUX by mass, MUX by mass, Min by mass, MUX (3) 158'5°C 0.05 0.01 99.0 0.06 0'03 10 5 To pass the test sulphate content ( on dry basis ), percent content ( on dry basis ), percent ( on dry basis ), percent a-phenylacetamide Phenylacetic Sulphated acid content ash, percent by mass, Max Lead ( as Pb ), ppm, MUX Arsenic ( as AssO, ), ppm, MUX Absence of cyanide APPENDIX ( Clause 2.2 ) METHODS A-l. QUALITY OF REAGENTS A OF TEST FOR a-PHENYLACETAMIDE material at 105 f 2°C. Cool and weigh. Repeat drying, cooling and till two subsequent weighings do not differ by more than O*OOlg. Reserve the dried material for tests under A-4 and A-5. A-3.2 Calculation Loss on drying, percent by mass = CM,where M1 = mass in g of the material the test, and M* = mass in g of dried material. A-4. DETERMINATION OF AMMONIUM SULPHATE CONTENT A-4.1 Outline of the Method - The material is extracted with water and then with ether. The water extract is treated with formaldehyde solution and then titrated with standard sodium hydroxide solution. From the amount of standard sodium hydroxide solution consumed, the total of ammonium sulphate present is quantity calculated. taken for M,) x 100 Ml A-l.1 Unless specified otherwise, pure chemicals and distilled water ( see IS : 1070-1977" ) shall be employed in tests. NOTE - `Pure chemicals' shall mean chemicals that do not contain impurities which affect the results of analysis. A-2. DETERMINATION OF MELTING POINT melting A-2.1 Procedure Determine the point according to the method prescribed in IS : 5762-1970t using the thermometer prescribed in A-2.1 .l. A-2.1.1 Thermometer - Thermometer shall of the mercury-in-glass type with a range from 85 to 215"C, graduated at every 0.2 deg, with partial immersion mark at 100 mm, and having a maximum error of f O-2 deg. A thermometer of other suitable range and accuracy may also be used. A-3. ;;~B.VIINATION OF LOSS ON A-3.1 Procedure - Weigh accurately about 5 g of the material in a tared porcelain dish and dry the *Specification for water for general laboratory use ( second revision ). tMethods for determination melting range. of melting point and 2 IS : 6393 - 1987 A-4.2 Reagents Ether See ZS : 336-1973". Solution See IS : A-5.1 Determination of Total Nitrogen A-4.2.1 A-4.2.2 Formaldehyde 3321-1973t. A-4.2.3 Standard Sodium Hydroxide Solution -- 0.01 N and 0.1 N. A-4.2.4 Phenolphthaiein O-5 g of phenolphthalem spirit ( see IS : 323-1959$ hydroxide solution until pink. Indicator - Dissolve in 100 ml of rectified ). Add standard sodium the indicator is faintly h-5.1.1 Apparatus - The apparatus, as assembled, is shown in Fig. 1. It consists of a roundhottom flask A, of 1000 ml capacity fitted with a rubber stopper through which passes one end of the connecting bulb tube B. The other end of the bulb tube B is connected to the condenser C by a rubber stopper and the lower end of the condenser C is attached by means of a rubber tubing to the dip tube D which dips into a known quantity of acid contained in a beaker E of 500 ml capacity, to which 3 to 4 drops of indicator soultion has been added. A-4.3 Procedure - Weigh accurately about 2 g of the dried material as reserved under A-3.1 in a separating funnel and to this add 30 ml of distilled water. Shake and add 30 ml of ether. Shake for three minutes. Allow to settle and remove the lower layer together with the precipitate in another separating funnel. Add ether three times ( 15 ml each time ) and extract. Collect all the ether extracts and wash with 20 ml of distilled water. Reserve tbe ether extract for test under A-6. Add water washings to the watery layer. Transfer water layer with precipitate to a conical flask and distil off ether, if any, on a water-bath. Cool and add 10 ml of formaldehyde solution previously neutralized with standard sodium hydroxide solution ( 0.01 N ) to light-pink colour using phenolphthalein indicator. Then titrate the solution with standard sodium hydroxide solution ( 0.1 N ) using phenolphthalein indicator. A-4.4 Calculation Ammonium sulphate, 6.6 V N percent by mass = M where V = volume in ml of standard sodium hydroxide solution, N -- normality of standard sodium hydroxide solution, and M = mass in g of the material taken for the test. A-5. DETERMINATION OF a-PHENYLACETAM-I-DE CONTENT Total nitrogen and nitrogen equivalent of ammonium sulphate Excess of total nitrogen is then are determined. calculated to equivalent quantity of a-phenylaceta_mide. A-5.0 Outline of the Method A-S.l.2 Reagents A-5.1.2.1 Potassium sulphate - anhydrous ( sodium sulphate may be used if potassium sulphate is not available ). A-5.1.2.2 Copper @hate A-5.1.2.3 Concentrated sulphuric acid IS : 266-l 977*. A-5.1.2.4 Standard sulphuric acid See 0.5 N. A-5.1.2.5 Methvl red indicator -- Dissolve 1 g of methyl red in 200 ml of rectified spirit ( see IS : 323-1959t ). A-5.1.2.6 Sodium hydroxide solution roximately 40 percent ( m/v ). tion - appsolu- A-5.1.2.7 Standard 0.25 N. sodium h)ldroxide *Specification for ether ( seconclrevision ). tspecification for formaldehyde solution ( fivsr revision ). A-5.1.3 Procedure - Weigh accurately about 0.5 g of the dried material as reserved under A-3.1 and transfer to a Kjeldahl flask. Add 10 g of powdered potassium sulphate and a few crystals of copper sulphate. Add 30 ml of concentrated sulphuric acid to the flask. Place the flask in an inclined position. Heat below the boiling point until frothing ceases. Raise the temperature to bring the acid to brisk boiling. Continue heating until the solution becomes straw-yellow in colour or practically water-white. Now remove the flask from the flame and cool. Transfer quantitatively to the round-bottom Bask A ( see Fig. 1 ) and dilute to about 250 ml with water. Add about 60 ml ( or more, if necessary, to make the solution alkaline ) of sodium hydroxide sclution carefully down the side of the flask so that it does not mix at -once with the acid solution but forms a laver below it. Assemble the apparatus as shown in Fig. 1 with the tip of the condenser dipping in a known quantity of standard sulphuric acid in beaker E to which a few drops of methyl red indicator have been added. Mix the contents of the flask by shaking and distil until all ammonia has passed over. Detach flask A from the condenser and shut off the burner. Rinse the condenser $Specification for rectified spirit ( revised ). 3 *Specificationfor sulphuric acid ( second revision ). tspecification for rectified spirit ( revised ). IS: 6393 - 1987 PIG. 1 DISTILLINGASSEMBLY FORDETERMINATION OF NITROGEN it4 = mass in g of the dried material taken thoroughly with water into the beaker E. Wash the for the test. dip tube D carefully so that all traces of the conWhen all densate are transferred to the beaker. A-5.2 Determination of Nitrogen in Ammonium the washings have drained into the beaker E, add Sulphate two or three drops more of the indicator and titrate with standard sodium hydroxide solution. A-5.2.1 Procedure - Carry out the test as prescribed under A-4.3. A-5.1.3.1 Carry out a blank using all reagents in the same quantites but without the material A-5.2.2 Calculation to be tested. Nitrogen in ammonium 1.400 V N A-5.1.4 Calculation sulphate, percent by mass = Total nitrogen, percent by mass = where VI = volume in ml of standard sodium hydroxide solution used to neutralize the acid in blank determination, -V, = volume in ml of standard sodium hydroxide solution used to neutralize the excess of acid in the test with the material, N = normality of standard sodium hydroxide solution, and 4 M 1.400 ( VI-V, A4 )N where V = volume in ml of standard sodium hydroxide solution ( see A-4.3 ), N = normality of standard sodium hydroxide solution ( see A-4.3 ), and M = mass in g of the material taken for the test ( see A-4.3 ). A-5.3 Determination of a-Phenylacetamide A-5.3.1 Calculation a-phenylactamide content, percent by mass = ( X1 X, ) x 9.654 Content IS : 6393 - 1987 where X, = percent by mass of total nitrogen A-5.1.4 ), and ( see A-7.2 Reagents A-7.2.1 Sulphuric Acid ( see IS : 266-1977" ). A-7.2.2 relative density 1.84 X2 = percent by mass of nitrogen in ammonium sulphate ( see A-5.2.2 ). .A& DETERMINATION ACID CONTENT A-6.1 Reagents A-6.1.1 Rectfj?ed Spirit See IS : 323-1959*. Nitric Acid - relative density 1.40 ( see IS : 264- 1976t ). A-7.3 Procedure -Weigh accurately about 1 to 2 g of the material in a tared crucible and ignite until thoroughly charred. Cool, moisten the residue with 1 ml of nitric acid and 1 ml of sulphuric acid, and cautiouslyignite until the carbon is completely consumed. Continue ignition in a place protected from air currents and using as low a temperature as possible until the combustion of the carbon is complete, cool the crucible in a desiccator and weigh. A-7.4 Calculation Sulphated ash, percent by mass = 100 ( Ml-M2 M ) OF PHENYLACETIC A-6.1.2 Phenolphthalein Indicator - Dissolve O-5 g of phenolphthalein in 100 ml of rectified spirit. Add standard sodium hydroxide solution until the indicator is faintly pink. A-6.1.3 Standard Sodium Hydroxide Solution 0.01 N. Transfer the ether extract as reserved under A-4.3 into a conical flask and evaporate the ether. Dissolve the residue in rectified spirit which was previously neutralized with the standard sodium hydroxide solution using phenolphthalein indicator. Titrate with standard sodium hydroxide solution using phenolphthalein indicator. A-6.3 Calculation Phenylacetic acid 13.6 V N content, percent by mass = M where v = volume in ml of standard sodium hydroxide solution, N = normality of standard sodium hydroxide solution; and M = mass in g of the dried material for the test ( see A-4.3 ). A-7. DETERMINATION ASH OF SULPHATED taken A-6.2 Procedure - where M, = mass in g of the ignited residue with the crucible, Mz = mass in g of the crucible, and M = mass~in g of the material taken for the test. OF LEAD ( AS Pb ) A-8. DETERMINATION A-8.0 Outline of the Method - The colour produced with hydrogen sulphide is compared to that produced by standard lead solution under identical conditions. A-8.1 Reagents A-8.1.1 Dilute Acetic Acid - 1:l, dilute acetic acid which complies with the following additional test. A-7.0 Outline of the Method-The material is ignited and burnt until only ash and carbon remain. After cooling, the charred residue is treated with nitric and sulphuric acids, and heated until oxidation of carbon is practically complete. The residue is then cooled and weighed. A-7.1 Apparatus A-7 .I .1 Crucible - squat form of silica or platinum. A-7.1.2 Bunsen Burner and Mufle Furnace *Specification for rectified spirit ( revised ). Evaporate 20 ml ~of the acid in a porcelain dish nearly to dryness on a&earn-bath. Add to the residue, 2 ml of acetic acid and dilute with water to 25 ml. Then add 10 ml of a solution of hydrogen sulphide. Any dark colour produced shall not be darker than a control made with 0.04 mg of lead and 2 ml of the diluted acetic acid ( 2 parts per million ). A-8.1.2 Hydrochloric Acid--Hydrochloric acid which complies with the following additional test. Evaporate 17 ml of the acid in a beaker to dryness on a steam-bath. Dissolve the residue in 2 ml of dilute acetic acid, dilute to 40 ml with water and add 10 ml of solution of hydrogen *Specificationfor sulphuric acid ( second revision ). tSpecification for nitric acid ( secondrevision). 1% 6393 - 1987 sulphide; any darkening produced shall not be greater than that in a blank test to which 0'02 mg of lead has been added ( 1 part per million ). A-8.1.3 Hydrogen SuIphide Solution prepared saturated solution of hydrogen in water. a freshly sulphide A-9. DETERMINATION OF ARSENIC ( AS As,% ) A-9.0 Outline of the~Mettrod - The stain produced by arsenic on a mercuric bromide paper is compared to stains produced by standard arsenic solution. A-9.1 Procedure-Weigh accurately I g of the material in 100 ml one-mark graduated flask ( see IS : 915-1975* ), add 25 ml of water and then sufficiently dilute hydrochloric acid ( 5 N ) to dissolve it. Make up the volume with water to 100 mI. Take 50 ml of this solution in a Gutzeit bottle and proceed as described in IS : 2088-19837 using for comparison 5 ml of standard arsenic trioxide solution, 1 ml of which contains O*OOl mg of arsenic trioxide (as Asz03 ). A-10. TEST FOR ABSENCE A-10.1 Reagents A-10.1.1 Ethanol - 50 percent. A-10.1.2 Sodium Hydroxide cent. Solution 10 perOF CYANIDE A-8.1.4 Lead Nitrate Stock Solution - Dissolve 159.8 mg of lead nitrate in 100 ml of water to which 1 ml of nitric acid has been added, then dilute to 1000 ml with water. NOTE-This solution shall be prepared and stored glass containers free from soluble lead salts. in A-8.1.5 Standard Lead Solution - Dilute 10 ml of the stock solution of lead nitrate accurately to 100 ml~with water. This solution shall be freshly prepared. Each millilitre of this standard lead solution contains the equivalent of 0'01 mg of lead. A-8.1.6 Test Solution a) Solutian A - Introduce into a 50-m] Nessler cylinder ( see IS : 4161-1967" ), 2 ml of dilute acetic acid and 1 ml of standard lead solution containing the lead equivalent of the heavy metals limit of 10 parts per million and make up to 25 ml with water. b) Solution B - Dissolve 1 g of the material under test in 10 ml of water, add 2 ml of dilute hydrochloric acid and water to make 25 ml. A-S.2 Procedure - Transfer solutions A and B to matching 50-m] Nessler cylinders, add 10 ml of solution of hydrogen sulphide to each tube, mix, allow to stand for 10 minutes, then view downwards over a white surface. A-8.2.1 The material shall be regarded to have not exceeded the limit prescribed if the colour intensity of solution B is not deeper than that of solution A. *Specificationfor Nessler cylinders. A-10.1.3 Hydrochloric Acid - Two parts of HCl in one part of water. A-10.1.4 Ferrous Sulphate A-10.1.5 Ferric Chloride A-10.2 Procedure -Take 0.5 g of sample in a 250 ml conical flask and dissolve in 10 nil of ethanol ( A-10.1.1 ). Add 2 ml of sodium hydroxide solution ( A-10.1.2 ) and 0.1 g of ferrous sulphate ( A-10.1.4 ). Boil the solution for 5 minutes. After cooling, add to the solution 5 ml of HCI ( A-10.1.3 ) followed by 0.5 ml of ferric chloride ( A-10.1.5 ). A-10.2.1 The material shall be -taken to be free from cyanide content if the colour of the solution does not become feeble green or blue. -*Specification revision revision ). for for one-mark volumetric of flasks (first tMethods ). determination arsenic ( second APPENDIX ( Clause 4. I ) B SAMPLE OF X+PHENYLACETAMIDE B-l. GENERAL B-l.1 Samples shall be taken at a place protected from damp air, dust and soot. 6 B-l.2 Sampling instrument shall be clean and dry. B-l.3 Precautions shall be taken to protect the sample, the material being sampled, the sampling IS:6393 - 1987 instrument and the containers for samples adventitious contamination. from B-2. SAMPLING INSTRUMENT B-l.4 To draw a representative sample, the contents of each container selected for sampling shall be mixed as thoroughly as possible by suitable means. B-l.5 The samples shall be placed in clean, dry and air-tight glass or other suitable containers on which the material has no action. B-l.6 The sample containers shall be of such a size that they are almost completely filled by the sample. B-l.7 Each sample container shall be sealed airtight after filling and marked with full details of sampling, the date of sampling, batch number and other important particulars of the consignment. B-l.8 Samples shall be stored in a cool and dry place. B-2.1 The sampling instrument is a closed type sampling tube, undivided ( see Fig. 2 ). It consists of two concentric cylindrical tubes made of a metal which is not affected by the material being sampled ( preferably of stainless steel ), one closely fitting into the other throughout their length so that it is possible to rotate one tube within the other, a suitable handle being provided for the purpose. Longitudinal openings of about one-third the circumference are cut in both tubes throughout their length. In one position, the two openings coincide and admit the material into the hollow inner tube. By rotating the inner tube through 180", the opening is tightly closed, a `core' of material being enclosed therein may be withdrawn. This type of sampler is usually provided with a locking arrangement so that the tubes are held The outer tube together in any desired position. is provided with a sharp conical-end to facilitate SECTION XX 9 ARRANGEMENT SECTION XX AFTER ROTATION OF INNER TUBE BY 180 CONE COMPLE TECY FIG. 2 CLOSED'_TYPE SAMPLINGTUBE, UNDIVIDED IS:6393 - 1987 B-3.2.1 These containers shall be selected at random from the lot and to ensure the randomness of selection, random number tables ( see IS : 4905-1968* ) shall be used. In case such tables are not available, the following procedure may be adopted: Starting from any container, count them in oneorderas 1,2,3 ,... , up to r and so on, where r is the integral part of N/n ( N being the lot size and it the number of containers to be selected ). Every ~rth container thus counted &all be withdrawn to give sample for test. B-4. TEST SAMPLE AND REFEREE SAMPLE B-4.1 From each of the containers selected as in B-3.2, draw with sampling instrument of an appropriate size, small portions of the material from different parts of the container. The total quantity so drawn from each of the containers shall be approximately equal to thrice the quantity required for testing purposes. B-4.2 Mix thoroughly all the portions of the material drawn from the same container to give a representative sample for the container. B-4.3 From the samples ( B-4.2 ) representing different containers selected in B-3.2, a small but equal quantity of material shall be taken and thoroughly mixed to form a composite sample sufficient to carry out testing for the characteristics specified. The composite sample so obtained shall be divided into three equal parts, one for the purchaser, another for the supplier and the third for the referee. B-4.4 The remaining portion of the material in the samples ( B-4.2 ) from different containers shall be divided into three equal parts, each forming an individual sample. One set of individual samples representing the n containers selected shall be for the purchaser, another for the supplier and the third for the referee. B-4.5 All the~individual and composite samples shall be transferred to separate containers. These containers shall then be sealed air-tight with stoppers and labelled with full identification particulars given in B-1.7. B-4.6 The referee samples, consisting of a composite sample and a set of n individual samples, shall bear the seals of both the purchaser and the supplier, and shall be kept at a place agreed to between the two. This shall be used in case of any dispute between the two. B-5. TESTS B-5.1 Tests for description ( see 2.1 ) and determination of melting point, and a-phenylacetamide content shall be conducted on each of the individual samples. *Methods for random sampling. penetration but the base of the cone shall be closed so that no material is entrapped in this portion. The height of the cone shall be equal to its base diameter. The whole instrument shall be of sufficient length to penetrate an entire diagonal of the container being sampled. The diameter of the inner cylindrical space may vary from 20 to 40 mm, proportionatelv to the length. A length of 150 cm and a diameter of 30 mm may cater for most needs. B-2.1.1 Use of Snmpling Instrument - The instrument is inserted in closed position in an oblique direction till it touches the bottom. The material is admitted by rotating and opening the tubes and finally closing them, with drawing the sample in this process. In case, the minimum quantity of material required for test from each container is more than the capacity of the instrument, further `cores' shall be taken from different parts of the same container such that they are at least 75 mm in the case of drums, bags, etc, and 25 mm in the case of small containers, from the wall of the container. In all cases, the instrument shall be inserted till it touches the bottom of the container so that an entire cross-section of its contents is withdrawn. B-3. SCALE OF SAMPLING B-3.1 Lot - All the containers in a single consignment of the material drawn from a single batch of manufacture shall constitute a lot. If a consignment is declared or known to consist of containers pertaining to different batches of manufacture, the containers belonging to the same batch of manufacture shall be grouped together and each such group shall constitute a separate lot. B-3.2 For ascertaining the conformity of the lot to the requirements of this specification, tests shall be carried out for each lot separately. The number of containers to be selected for drawing the samples shall depend on the size of the lot and shall be in accordance with Table 2. TABLE 2 NUMBER OF CONTATNERS TO BE SELECTED FROM LOT OF DIFFERENT SIZES LOT (N) 4 to 2.5 26,,50 51 ,, 100 101 ,, 150 151 ,, 300 301 and above SIZE No. OP CONTAINERS To BE SELECTED (h) 3 4 5 6 7 8 or less, all NOTE - When the size of the lot is three the containers shall be sampled. 8 IS : 6393 - 1987 B-5.2 Tests for determination teristics shall be conducted sample. of all other characon the composite content if each of the test results satisfies the corresponding requirements given in 2.1 and Table 1. B-6.2 For Composite Sample - For declaring the conformity of a lot to the requirements of all other characteristics tested on the composite sample ( see B-4.2 ), the test results shall satisfy all other requirements specified in this standard. B-6. CRITERIA FOR CONFORMITY B-6.1 For Individual Samples - The lot shall be declared as conforming to the requirements of description, melting point and a-phenylacetamide